Efficient conditional transgene expression in hepatitis C virus cDNA transgenic mice mediated by the Cre/loxP system
Efficient conditional transgene expression in hepatitis C virus cDNA transgenic mice mediated by the Cre/loxP system
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DOI：
10.1074/jbc.273.15.9001
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发表时间：
1998-04-10
影响因子：
4.8
通讯作者：
Kohara, M
中科院分区：
文献类型：
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作者：
Wakita, T;Taya, C;Kohara, M
条件性基因表达极大地方便了对特定基因产物功能的研究，利用Cre/loxP系统，我们在转基因小鼠中建立了高效的条件转基因激活丙型肝炎病毒(HCV)基因(294-3435核苷酸)，静脉注射表达Cre DNA重组酶的腺病毒，在转基因小鼠的肝脏中观察到了有效的重组。转基因激活后，大多数肝细胞被抗核心多克隆抗体染色，用抗核心、EL和E2单抗分别在肝裂解液中检测到21、37和蛋白，转基因小鼠在激活后7天检测到血清核心蛋白，同时血清丙氨酸氨基转移酶水平升高，随后在感染14天后检测到抗核心抗体应答，并进行了CD_4和CD_8阳性细胞清除试验，使血清丙氨酸转氨酶升高和肝脏病理改变正常化。这些结果表明，丙型肝炎病毒蛋白不是直接的细胞病变，宿主免疫应答在丙型肝炎病毒感染中起着关键作用，因此，该转基因小鼠为研究丙型肝炎病毒感染的免疫应答和致病机制提供了有力的工具。
Conditional gene expression has greatly facilitated the examination of the functions of particular gene products, Using the Cre/loxP system, we developed efficient conditional transgene activation of hepatitis C virus (HCV) cDNA (nucleotides 294-3435) in transgenic mice, Efficient recombination was observed in transgenic mouse liver upon intravenous administration of adenovirus that expresses Cre DNA recombinase. After transgene activation, most hepatocytes were stained with anti-core polyclonal antibody, and 21-, 37-, and 64-kDa proteins were detected by Western blot analysis in liver lysates using anti-core, El, and E2 monoclonal antibodies, respectively, Serum core protein was detected in transgenic mice 7 days after transgene activation with concurrent increases in serum alanine aminotransferase levels, Subsequently, an anti-core antibody response was detected 14 days after infection, Furthermore, a CD4 and CD8 positive cell depletion assay normalized both the serum alanine aminotransferase increases and pathological changes in the liver, These results suggest that HCV proteins are not directly cytopathic and that the host immune response plays a pivotal role in HCV infection, Thus, this HCV cDNA transgenic mouse provides a powerful tool with which to investigate the immune responses and pathogenesis of HCV infection.